How to prevent denaturation of fragile biomolecules and guarantee their proper functioning after several weeks of storage time typical for products on the market?
Dear prof. Camman. I am working on the same problem in the field of metal oxide nanostructures for biosensor applications. Can we discuss it via skype or e-mail? Sincerely, Roman
What about a storage in glycine buffer at a slight alkaline pH value?
Dear Karl the answer is easy!
Just dont use fragile molecules but sturdy molecules ... like aptamers !
oligonucleotide aptamers or peptide aptamers !
Dear Marco,
thanks for your suggestion. I have witnessed too many failures of aptamers during my time as mid-term reviewer of the European Commission (the same with molecular imprinted polymers – MIP). Can you send me a link of a company which sells pre-calibrated sensors with aptamers. Up to know I don’t trust the expectations: better stability and at the same time excellence specifity; but if they are really successful devices with aptamers on the market, I will change my mind immediately!
Best regards
Karl
Dear Karl there is a big difference with MIP.
MIP are just a kitchen recipe ... there is not rational at all.
With aptamers we start from a huge amount of compounds and we look for a specific good ligand ! the cycle is very important (SELEX and similar!) .
For a Company what about Affibody from Sweden ?
Very good material !
Best wishes for the coming Holidays ! Marco
Dear Karl,
When fragile molecules are regenerated he Rmax value for analyte binding is changing and after a few regeneration steps the response is totally gone. Definitely it is necessary to start always with an equal quality of sensor array. Best is to capture the fragile molecules using a tag by having anti-tag antibodies on the surface.. (Or if not possible a relevant epitope should be captured) using anti-epitope antibodies. There are several very high quality anti-tag antibodies available in the market including Myc, Flag, His, etc... tags . So a two step interaction should take place. First capture using the tags and record the signals followed by the multiplex analyte broth. The microarray sensor array can be stored and the quality can always be checked with the captured signal of the ligand molecules using label-free SPR imaging e.g. the IBIS MX96 instrument for multiplex label free interaction. In the interaction lab of IBIS technologies there is great expertise for fabricating the microarray using continuous flow microspotting. www.ibis-spr.nl Thank you.....
Dear Prof. Karl Cammann
By means of this communication, I suggest the use of microarrays (MEMS) based on aptamers which are described briefly in my chapter about of biosensors found in the next web page: http://www.intechopen.com/books/pesticides-advances-in-chemical-and-botanical-pesticides/evolution-and-expectations-of-enzymatic-biosensors-for-pesticides. Important technical details must be considered as the previous comments have introduced. Dr. Rafael Vargas Bernal.
Many thanks for all the valuable suggestions. I am looking for a successful biosensor on the market (besides glucose with the exceptionally stable glucose oxidase) which survived a thorough validation study according to the ISO norms. That is demonstrated in an inter-laboratory comparison study that the calibration at the production factory resulted in true results at the customer, independent of the biomolecule (enzyme, antibody or aptamers). I could not find a single biosensor still sold on the market which was also tolerated by accreditation bodies. I am looking for a bit more than nice cartoons how it might work or had worked in a research lab.
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